CN107064487A - A kind of graphene accelerates elisa technique - Google Patents
A kind of graphene accelerates elisa technique Download PDFInfo
- Publication number
- CN107064487A CN107064487A CN201710077713.1A CN201710077713A CN107064487A CN 107064487 A CN107064487 A CN 107064487A CN 201710077713 A CN201710077713 A CN 201710077713A CN 107064487 A CN107064487 A CN 107064487A
- Authority
- CN
- China
- Prior art keywords
- elisa
- antibody
- antigen
- graphene
- accelerates
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
Abstract
" a kind of graphene acceleration elisa technique " of the invention, it is characterized in that a kind of new material graphene accelerates the improved ELISA technology that antigen-antibody is combined with the ELISA new capabilities being combined, utilize the absorption of graphene height and high-ratio surface characteristic, increase the contacting efficiency between antigen-antibody in ELISA reaction systems, accelerate ELISA (enzyme linked immunosorbent assay (ELISA)) reactions and coating process.The technology is related to immunology and field of immunodetection, it is the theory innovation that a kind of graphene is combined with ELISA, using graphene there is binding molecule and high-ratio surface characteristic to increase the contact probability between antigen-antibody, the problem of solving ELISA reactions and coating overlong time.
Description
Technical field：
The invention belongs to immunology and field of immunodetection, and in particular to the high absorption and high-ratio surface by graphene are special
Property increase the contact between antigen and antibody, accelerate the technical field of ELISA (enzyme-linked immunosorbent assay) course of reaction.
Background technology：
An immune word is developed by Latin word " immunis ", and its original meaning is " release tax revenue "
(exceptionfromcharges) meaning of " suffering from from epidemic disease ", is also included.Immune narrow sense concept in life science refers to body
" itself " and " non-oneself " antigen are recognized, to autoantigen formation natural immune tolerance, repulsive interaction is produced to " non-oneself " antigen
A kind of physiological function.Under normal circumstances, this physiological function is beneficial to body, can produce the maintenance body such as anti-infective, antitumor
Physiological equilibrium and stable immanoprotection action.Under certain condition, when immune dysfunction, body can also be produced harmful
Reaction and result, such as trigger hypersensitivity, autoimmunity disease and tumour.
Immunology is to study organism confrontation original matter immunologic responsiveness and its biology-medical science of method.It is immune to answer
Answer be body fight primary stimuli reaction, be also a kind of biological process that antigenic substance is identified and excluded.Typically recognize
For immunologic development experience four periods be experience immunology period, classical immunology period, modern age immunology period and
Immunology Today period.
1st, experience immunology period：Early in 11st century of Christian era, Chinese medicine man has creatively invented people's acne in practice
Seedling, i.e., manually the method for low-grade infection prevents smallpox.Between the Ming Dynasty's grand celebrating year (1567~1572), humanized lymph is Chinese wide
General application.Cause the attentions of other countries in the inoculate against method of smallpox of the 17th century humanized lymph, successively incoming Russia, Korea,
The ground such as Japan, Turkey, Britain, and then enable humanized lymph to prevent the method for smallpox to promote and verify that this is experience immunology
Period, the period is found that immunological phenomena, is the beginning of human knowledge's immunity of organism, and British physician Jenner is invented after being
Bovine vaccine is laid a good foundation.
2nd, experience immunology period：18th century to 20 middle of century are classical immunology period, in this period, people couple
The understanding of immunologic function enters scientific experiment period by the observation of human body phenomenon.First, the invention of bovine vaccine is British physician
Jenner is observing the milkmaid of suffering from cowpox, after the fact that no longer suffer from smallpox, passes through the scientific payoffs studied for a long period of time.Ox
The invention of bovine vaccine is an immunologic important development after humanized lymph, after the vaccine is to human vaccination, only causes local anti-
It should not cause serious harm, but can effectively prevent smallpox, also can largely be produced in laboratory, therefore soon instead of people
Bovine vaccine, is received by medical field.Secondly, the invention of attenuated live vaccine is French immunologist Pasteur and German bacteriologist
Koch, by systematically scientific research, utilizes physics, chemistry, biology on the basis of bacteria distribution culture technique has been founded
Method obtains attenuation vaccine, and for prevention and treatment of diseases, Pasteur is prepared for Anthrax vaccine with high-temperature cultivation method,
With rabies viruses rabies vacciness is prepared in rabbit body through continuous passage.Again, antibody is that Behring and Kitasato exist
Found when animal is immunized with diphtherotoxin within 1890.There is one kind to neutralize ectotoxic thing in by immune animal blood serum
Matter, is initially known as antitoxin.By this immune serum passive transfer to intact animal, the latter is set to obtain the ectotoxic energy of neutralization
Power.Diphtheria antitoxin (DAT) is formally used for the treatment of diphtheria by above-mentioned scientist, has started the beginning of artificial immunity therapy, therefore
Behring obtained Nobel's medical science and physiology prize in 1901.Later, people were found that the energy such as agglutinin, precipitin in succession
The material reacted with bacterium or cell-specific, is referred to as antibody；And the material referred to as antigen that antibody will can be caused to produce, so that
Establish the concept of antigen and antibody.Then it is found that immune bacteriolysis in Pfeiffer in 1894 and then is found that complement,
Complement is a kind of material to thermally labile, the composition belonged in normal serum, no specificity, but with assisting antibody dissolving thin
Bacterium or the effect of cell.With the theoretical proposition of the foundation, immunochemical research, antibody tormation of serological method and to machine
The research of body protective immunity mechanism, immunologic development accelerates speed.
3rd, modern age immunology period：It it is immunology period in modern age during 20 middle of century to the sixties.This period people
The constraint of anti-infectious immunity template theory is broken through, the immunoreactivity to organism has than more comprehensively understanding, makes to be immunized
Begin one's study biological questions, occurs in that brand-new Immunology.This period is found that delayed allergy, immune resistance to
By phenomenon, it is proposed that clonal selection theory, immunological technique is rapidly developed, indirect agglutination and immune mark are established
Note technology, these all further promote the theoretical research and application of amynologic basis.
4th, modern age immunology period：Immunology Today period refers to the period of 1960s so far.In this period, really
The status that lymphocyte of accepting is tied up in immune response, illustrates the molecular structure and function of immunoglobulin, to immune system
Particularly cell factor, adhesion molecule etc. has carried out numerous studies, and from diversity of the molecular level to immunoglobulin, classification
Conversion etc. has carried out beneficial discussion, and breakthrough achievement is achieved in many aspects.Applied immunology field, 1975
Kohler and Milstein initiates hybridoma technology.They are by murine myeloma cell and through sheep red blood cell (SRBC) (SRBC) sensitization
B cell carries out fusion and forms hybridoma (hybridoma) in vitro.It is a large amount of that this hybridoma had both maintained myeloma cell
The characteristic of unrestricted growth and breeding, has the ability of synthesis and secretory antibody again.Using the technology can produce it is homogeneous, just for
The antibody of single epitope, referred to as monoclonal antibody (McAb).McAb has purity height, high specificity, can largely produced
The advantages of, be widely used in the detection of serodiagnosis, immunocyte and other histocyte surface moleculars, and by with
Nucleic, various toxin or pharmaceutical chemistry coupling carry out tumor targeting therapy research.Protocols in Molecular Biology is applied to immunology
Research is also a breakthrough achievement, and the genetic engineering antibody prepared using molecular hybridization and molecular genetic theory is for example complete
Full humanized antibody, single-chain antibody and bispecific antibody etc. have more superiority compared with McAb.
Antigen (antigen, Ag) be it is a kind of can cause the material of specific immune response in organism, the material enters
Body can be stimulated to produce after body and antibody and cause cellular immunity, in immunoassays antigen refer to can be with antibody binding thing
Matter.The reactionogenicity of so-called antigen refers to can occur specificity instead with the antibody produced by being stimulated by it or sensitized lymphocyte
Should, possess the material referred to as comlete antigen of two kinds of abilities of immunogenicity and reactionogenicity, such as pathogen, heterogenous animal serum.
Only there is reactionogenicity without the material of immunogenicity, referred to as haptens, without immunogenicity, immune response will not be caused,
Such as penicillin, sulfanilamide (SN).But it is some in particular cases, if after haptens is combined with macro-molecular protein, just exempted from
Epidemic focus and become comlete antigen, also just antibody and effector cell can be produced with stimulating immune system.The fundamental property tool of antigen
There are foreign body, macromoleculariness and specificity.Specifically, the foreign body antigenic substance for referring to enter in body tissue, it is necessary to
The composition of the body tissue cell is differed；The material that macromoleculariness refers to constitute antigen is typically that relative molecular mass is more than
10000 macromolecular substances, molecular weight is bigger, and antigenicity is stronger；Specificity refer to a kind of antigen can only with corresponding antibody or
Effector T cell is specifically bound.Two classes are divided into according to antigenic property：Comlete antigen and incomplete antigen.Comlete antigen
(complete antigen) abbreviation antigen, is the existing immunogenicity of a class, there is immunoreactive material again, such as most of eggs
White matter, bacterium, virus, bacterial exotoxin etc..Incomplete antigen is also known as haptens (hapten), is only with immunoreactivity
The material of non-immunogenicity, after haptens is combined with protein carrier, just obtains immunogenicity.
Antibody (antibody) is a kind of to be used for differentiating and neutralized by immune system by what thick liquid cell (effect B cell) was secreted
The large-scale Y shape protein of foreign substance such as bacterium, virus etc., narrow sense concept refers to the immune globulin that can be combined with antigentic specificity
In vain (immunoglobulin, Ig), the predominantly IgG and IgM relevant with ELISA.Antibody has the symmetrical junction of 4 polypeptide chains
Structure, wherein the identical heavy chain (H chains) 2 longer, relative molecular weight is larger, 2 shorter, the less identical of relative molecular weight
Light chain (L chains), interchain is coupled by disulfide bond and non-covalent bond forms a monomer molecule being made up of 4 polypeptide chains.Light chain has κ
With two kinds of λ, heavy chain has five kinds of μ, δ, γ, ε and α, and whole antibody molecule can be divided into constant region and variable region two parts.What is given
In species, the constant region of different antibodies molecule all has identical or almost identical amino acid sequence, and variable region is located at " Y "
Two-arm end, has the change of sub-fraction amino acid residue especially strong in variable region, the residue composition of these amino acid and row
The row order region that is more easy to morph claims hypervariable region, and hypervariable region is located at molecular surface, is at most made up of 17 amino acid residues, few
Then there was only 2~3, hypervariable region amino acid sequence determines the specificity of the antibodies bind antigen antigen.On one antibody molecule
Two antigen-binding sites be identical, claim antigen-binding fragment (antigen-binding positioned at two-arm end
Fragment, Fab), the shank of " Y " claims crystallizable fragment (crystalline fragment, FC), and sugar is combined on FC.According to reason
Change property and biological function, antibody can be classified as the class of IgM, IgG, IgA, IgE, IgD five.The preparation that Ankang is carried, antibody can
It is divided into monoclonal antibody and polyclonal antibody.The main function of antibody is combined with antigen (including external and itself),
So as to effectively remove the foreign matters such as microorganism, parasite in intrusion body, antibody (antibody) is a kind of response antigen production
Protein that is raw, being combined with antigentic specificity, every kind of antibody is combined with specific epitope, and this combination can make
Antigen is inactivated, it is also possible to invalid but also can cause pathologic lesion to body sometimes.
It is to be combined by very weak short distance gravitation between antigen-antibody, the adhesion is a kind of the non-of high degree of specificity
Covalent bond effect power, such as hydrogen bond, electrostatic attraction, hydrophobic interaction power, Van der Waals force.Wherein hydrophobic interaction power and
Electrostatic attraction is main active force, and only both active forces make antibody and antigen molecule close to a certain extent, Van der Waals
Power and hydrogen bond action can just be functioned to.Several active forces are weaker than covalent bond effect above, only exist substantial amounts of above-mentioned
There is extraordinary cooperation between active force, and antigen and antibody, useful effect could occur between antigen and antibody, this has reacted anti-
The specific feature of original antibody wording depth.The affinity of antibody refers to the intrinsic adhesion between antigen-antibody, can use balance
Constant K is represented：K=[AgAb]/[Ag] [Ab] AgAb dissociation degree is relevant with K values, the antigen position of high-affinity antibody
Point and the determinant of antigen are most suitable in steric configuration, and the two is firmly combined with, and is difficult to dissociate.Antigen-antibody reaction has most
Suitable ratio, this most suitable scope is referred to as equivalence zone, is respectively antibody excess zone and zone of antigen excess before and after equivalence zone.If antigen
Or antibody is extremely superfluous, then deposit-free is formed, and zoning is referred to as in immunoassays, and antibody excess is referred to as preceding band, antigen mistake
It is surplus to be referred to as rear band.When with determination of immunological methods antigen, it should make have enough antibody levels in reaction system, otherwise measure
Amount can be less than actual content, or even false negative occur.Antigen-antibody reaction has specificity, and the combination of antigen-antibody is substantially only
Occur between the antigenic determinant of antigen and the antigen binding site of antibody, because the two is in chemical constitution and steric configuration
In complementary relationship, so antigen-antibody reaction has high degree of specificity.Antigen-antibody reaction also has sensitiveness, is determining serum
When middle content of material, the susceptibility that enzyme reaction is determined is about 5-10 μ g/ml, and chemical colorimetry sensitivity is mg/ml water
It is flat.
Enzyme linked immunosorbent assay (ELISA) (Enzyme Linked Immunosorbent Assay, ELISA) refers to solubility
Antigen or antibody binding on the solid phase carriers such as polystyrene, carry out enzyme labeled immunoassay using antigen-antibody binding specificity anti-
The qualitative and quantitative detecting method answered.Engvall and Perlmann in 1971, which has delivered enzyme-linked immunosorbent assay, is used for IgG
The article of quantitative determination, its general principle is：1. antigen or antibody is physical is attached to certain surface of solid phase carriers, and keep it
Immunocompetence.2. antigen or antibody connect into enzyme-labelled antigen or antibody with certain enzyme, and this enzyme-labelled antigen or antibody both retain it
Immunocompetence, retains the activity of enzyme again.3. being pressed by inspection sample (determine therein antibody or antigen) and enzyme-labelled antigen or antibody
Different steps and the antigen of surface of solid phase carriers or antibody are reacted, and the antigen formed on solid phase carrier is made with the method for washing
Antibody complex is separated with other materials, finally with reference to tested substance in the enzyme amount and sample on solid phase carrier amount into certain
Ratio, add enzyme reaction substrate after, substrate is changed into tested substance in color products, the amount and sample of product by enzymatic
Amount is directly related, therefore can be according to the depth qualitative or quantitative analysis of color reaction.Because the catalytic efficiency of enzyme is very high, therefore can be very big
Ground iodine effect, so that assay method reaches very high susceptibility.
ELISA gets up with immunologic development, be widely used to protein engineering, medical treatment, agricultural,
The series such as herding, environmental protection, food field, the Protein Detection field especially in disease treatment, disease detection, scientific research is obtained extensively
Using having become the important technology of 21st century life science.Its main method has following four kinds：
1st, Salmonella：It is, with coating buffer solution dilution coating to solid phase carrier, to wrap antigen according to certain ratio
Simply washed after being done, add confining liquid closing, washed away more than confining liquid, add appropriate enzyme-specific antibody, 37 DEG C
1h or 4 DEG C of overnight incubation is incubated, washing removes unnecessary antibody, and result, the colour developing depth and addition are read in addition substrate colour developing
Enzyme labelled antibody amount is directly proportional.The method is primarily adapted for use in detection monoclonal antibody hypotype, serum kind and primary dcreening operation monoclonal antibody.
2nd, indirect method：Its principle be using enzyme mark antiantibody detection combined with solid phase antigen by examine antibody, be by
Antigen, with coating buffer solution dilution coating to solid phase carrier, is simply washed after the completion of coating, added according to certain ratio
Confining liquid is closed, and washes away unnecessary confining liquid, adds appropriate non-enzyme labelled antibody, is introduced by second of antibody of enzyme target (with the
A kind of antibody of antibody binding, i.e. secondary antibody) with the first antibody (with antigen directly in conjunction with antibody, i.e. primary antibody) specificity knot
Close, be eventually adding substrate colour developing and judged result, when the concentration of secondary antibody is certain, the amount of colour developing result and primary antibody is in positive
Close.Indirect ELISA is the important means for detecting significant antibody, mainly for detection of pathogen antigen, antibody titer, serum effect
Valency, monoclonal antibody etc., general operating procedure is：1. known antigen is bonded into solid phase carrier to be coupled, it is unnecessary to wash away
Uncombined antigen and impurity.2. the specific antibody added in sample to be tested, sample to be checked is combined with solid phase antigen, forms solid phase
Other compositions are left behind in specific antibody, sample to be checked on antigen antibody complex, scrubbed rear solid phase carrier washed
Washed away in journey.3. add the antibody carried in enzyme target secondary antibody, solid-phase immunity compound to be combined with enzyme labelled antibody, so that indirectly
Mark on enzyme.4. unnecessary uncombined secondary antibody is washed away, substrate colour developing is added and reads result, the colour developing depth resists with the enzyme mark added
The scale of construction is directly proportional.
3rd, sandwich ELISA：Be otherwise known as sandwich ELISA, and the method is usually used in detecting macromolecular antigen, can be divided into direct folder
Heart ELISA and indirect sandwich ELISA, the former is divided into double crush syndrome and double antigens sandwich ELISA again.Direct double antibodies sandwich
ELISA is detection antigen most common method, and its cardinal principle is：The first antibody is coated on solid phase carrier, after closing
Antigen to be checked is added, is added after incubation and passes through enzyme second of antibody of target, the first antibody and second of antibody can be directed to
Two kinds of monoclonal antibodies of different epitopes or a kind of monoclonal antibody for same antigen resist more with a kind of.Its operating procedure is as follows：1. will
Specific antibody is coupled with solid phase carrier, forms insolubilized antibody, washes away uncombined antibody and impurity.2. sample to be tested is added,
Antigen in sample to be checked is combined with solid phase antigen, forms solid phase antigen antibody complex, is washed to remove other uncombined things
Matter.3. add the antigen carried in enzyme target secondary antibody, solid-phase immunity compound to be combined with enzyme labelled antibody, be washed to remove other not
Enzyme amount on conjugate, solid phase carrier by the amount of inspection antigen with being proportionate.4. add substrate colour developing and read result, colour developing is deep
It is shallow to be directly proportional to adding by inspection antigen.
4th, competitive ELISA：Also known as Competitive assays ELISA or closing ELISA, its principle is to go to do with antigen to be checked or antibody
Pre-designed system is disturbed, the annoyance level of the result finally developed the color and antigen to be checked or antibody, into negative correlation, is one
The less detection method used is planted, is generally used for detecting small molecule antigens, its operating procedure is：1. by the antibody with selectivity
It is bonded on solid phase carrier, washes away Excess antibody.2. sample to be tested is added, the determined antigen in sample is carried out with insolubilized antibody
Selectivity is combined.3. the antigen marked with enzyme is added, this antigen also can carry out selectivity with insolubilized antibody and be combined, due to solid phase
Antibody levels are limited, therefore when the amount of antigen in sample is more, then the antigen and insolubilized antibody marked with enzyme is fewer, that is,
Two kinds of antigens are all competitively combined with insolubilized antibody.4. sample and the antigen marked with enzyme are washed away, substrate colour developing is added, works as sample
Middle amount of antigen is more, and the antigen marked with enzyme is fewer, develops the color also more shallow, and colour generation result is read with naked eyes or instrument.
Graphene (Graphene) is the extremely important new material found over year more than last decade, graphene narrow sense concept
Refer to the two dimension crystalline substance for being made up of the only one of which carbon atom thickness of hexangle type honeycomb lattice structure with sp2 hybridized orbits carbon atom
Body.The graphene number of plies is in normal distribution, and most of broad sense graphene refers to multi-layer graphene, such as form the few-layer graphene alkene (3~5 layers),
Multi-layer graphene (10 layers or so).Univ Manchester UK physicist An Deliehaimu and Constantine Nuo Woxiaoluo
Husband isolated graphene in 2004 from graphite, and confirmed that graphene can be with individualism, and two people are also because in " two-dimentional graphite
The pionerring research of alkene material " obtains Nobel Prize in physics in 2010 jointly.Graphene is good conductor, not only very hard
Hard and electric conductivity is splendid, its electron mobility is up to 200,000cm2V-1s-1, resistivity is up to 10-6Ω cm, are in the world
The minimum material of resistivity, is also splendid normal temperature superconductor.Graphene also has preferable bio-compatibility and absorption each
Plant the characteristic of atom and molecule.
Antigen and antibody belong to protein molecule, extremely small, and the relative distance between its molecule is larger, each other from
Farther out, if it is possible to increase the contact between antigen and antibody, it becomes possible to increase the efficiency of antigen and antibody binding.Due to graphite
Alkene has preferable bio-compatibility, while having the various atoms of absorption and molecule and up to 2600m2/ g than surface characteristic,
The adsorbable antigen of graphene and (or) antibody, and be further dispersed to rapidly in the space of antigen-antibody combination, greatly accelerate anti-
Contact area and joint efficiency between original antibody.And conventional anti-ELISA is just with non-covalent bond effect power, such as hydrogen bond, quiet
Electric attraction, hydrophobic interaction power, Van der Waals force, by graphene and antigen, the combination of antibody, greatly shorten the two
Binding time, it is ensured that the reliability of result.
The content of the invention：
The problem of in order to solve ELISA reaction time and coating overlong time, " a kind of graphene accelerates ELISA to the present invention
Technology " combines a kind of improved ELISA technology of new material graphene with the ELISA acceleration ELISA processes being combined, and utilizes stone
Black alkene has the contact area and efficiency between the characteristic of high absorption and high-ratio surface, increase antigen-antibody, substantially reduces
In the ELISA reaction time, while coating process can be accelerated, allow scientific research and clinical examination personnel to understand testing result in time, improve
Detection efficiency and accuracy.
A kind of graphene accelerates elisa technique, it is characterized in that the new capability that a kind of new material graphene is combined with ELISA
The improved ELISA technology for accelerating antigen-antibody to combine, using the absorption of graphene height and high-ratio surface characteristic, in ELISA reactants
Increase the contact between antigen-antibody in system, accelerate ELISA (enzyme linked immunosorbent assay (ELISA)) to react and coating process.Above-mentioned institute
The ELISA stated refers to, by soluble antigen or antibody binding to the solid phase carriers such as polystyrene, combine using antigen-antibody
Selectivity carries out the qualitative and quantitative detecting method of immune response.
A kind of graphene accelerates elisa technique, it is characterised in that described new material graphene narrow sense concept refers to by carbon
Atom constitutes the two dimensional crystal of the only one of which carbon atom thickness of hexangle type honeycomb lattice structure with sp2 hybridized orbits, can also adopt
It is preferred especially with the multi-layer graphene within 10 layers with broad sense graphene (multilayer), and with higher absorption and high-ratio surface characteristic
Graphene.
A kind of graphene accelerates elisa technique, it is characterised in that its English of described ELISA is expressed as Enzyme
Linked Immunosorbent Assay, its Chinese is expressed as enzyme linked immunosorbent assay (ELISA), refer to soluble antigen or
Antibody binding utilizes antigen-antibody binding specificity to carry out the qualitative or quantitative of immune response on the solid phase carriers such as polystyrene
The process of detection.
A kind of graphene accelerates elisa technique, it is characterised in that described graphene adds with the ELISA new capabilities being combined
The improved ELISA technology that fast antigen-antibody is combined, can be appropriate according to the addition of specific substance characteristics before, during and after ELISA processes
Graphene, to shorten the ELISA reaction time, solve ELISA one detection periods long (needing 2~3 hours) and coating
The problem of overlong time (needing to stay overnight), make to understand experiment or testing result within a short period of time and obtain more reliable data
It is possibly realized.
A kind of graphene accelerates elisa technique, it is characterised in that described antigen is that one kind can cause spy in organism
The material of specific immunological response, the material, which enters, can stimulate body to produce antibody and cause cellular immunity after body, in immune survey
Fixed middle antigen refers to material that can be with antibody binding；It is characterized in that described antibody be one kind by thick liquid cell (effect B cell)
The large-scale Y shape protein for being used for differentiating and neutralizing foreign substance such as bacterium, virus etc. by immune system of secretion.
A kind of graphene accelerates elisa technique, it is characterised in that described antigen-antibody, which is combined, refers to that antigen has with antibody
There is the characteristic that selectivity is combined, i.e. its corresponding antibody of antigen has specificity structure, combined by very weak short distance gravitation
, the adhesion is a kind of non-covalent bond effect power of high degree of specificity, can be specifically bound to a certain extent.
A kind of graphene accelerates elisa technique, it is characterised in that the contact between described increase antigen-antibody refers to lead to
Graphene absorption, scattered antigen and (or) antibody molecule are crossed, to increase the contact probability between antigen and antibody, and then improves anti-
The former joint efficiency with antibody, can not only accelerate to react (incubations) process, can also accelerate coating process, it is above-mentioned it is tactful not only can be with
For conventional ELISA, also available for fluorescence elisa technique, chemiluminescence elisa technique, PCR-ELISA technologies, NSP-
Elisa technique, Dot-ELISA technologies, TP-ELISA technologies, IMS-ELISA technologies, PPA-ELISA technologies etc..
A kind of graphene accelerates elisa technique, it is characterised in that described graphene is combined increase antigen with ELISA and resisted
Contact probability this strategy between body can greatly reduce ELISA reactions and coating time, it is adaptable to herding, agricultural, food peace
Entirely, clinical diagnosis, scientific research detection and enzyme immune detection and the field of immunology of diagnosis, also suitable for external diagnosis reagent II classes
With the immune related reagent in III products, apply also for detecting antigen, antibody and the haptens etc. in other samples.
A kind of graphene accelerates elisa technique, it is characterised in that high absorption and high ratio by a kind of new material graphene
Surface characteristic accelerates ELISA strategy to be applied to computer software programs and network journey come the contact increased between antigen and antibody
Sequence is write, and is improved ELISA detection efficiencies, reduction ELISA reactions and coating time, is further improved graphene and tied with ELISA phases
The immunological technique of the acceleration ELISA processes of conjunction.
A kind of graphene accelerates elisa technique, it is characterised in that high absorption and high ratio by a kind of new material graphene
Surface characteristic accelerates ELISA strategy to be applied to ELIASA, fluorescence microplate reader, light splitting come the contact increased between antigen and antibody
Photometer, sepectrophotofluorometer, automatic enzyme non-analysis meter etc. provide the Instrument Design of detection absorbance and develop to improve
ELISA reacts and coated efficiency, and its main component includes coating plate, enzymic-labelled antibody and (or) antigen, standard items, buffering
Liquid, nitrite ion, terminate liquid, cleaning solution etc..
" a kind of graphene acceleration elisa technique " purpose of the invention is to add in order to increase the contact between antigen and antibody
Fast ELISA (enzyme linked immunosorbent assay (ELISA)), reaction time and coating time, which shorten, to be more beneficial for observation result and ensures that its is reliable
Property.ELISA basis is the immobilization and antigen or the enzyme mark of antibody of antigen or antibody, with reference in the anti-of surface of solid phase carriers
Former or antibody remains in that its immunologic competence, and the antigen or antibody of enzyme mark had not only retained its immunologic competence but also retained the work of enzyme
Property.ELISA basic steps are as follows：1. antigen or antibody binding is to surface of solid phase carriers and keeps its immunocompetence；2. antigen or
Antibody connects into enzyme-labelled antigen or antibody with certain enzyme, and the enzyme-labelled antigen or the existing immunocompetence of antibody have enzymatic activity again；3. treat
Examine sample (determining antibody therein or antigen) and enzyme-labelled antigen or antibody presses the antigen of different step and surface of solid phase carriers
Or antibody reacts, uncombined antigen or antibody and other materials is washed away, with reference in the enzyme amount on solid phase carrier and sample
Amount of substance to be checked adds substrate after the substrate of enzyme reaction and is changed into color products, product amount and mark by enzymatic into certain ratio
Amount of substance to be checked is directly related in this, therefore can be according to the depth qualitative or quantitative analysis of color reaction.Due to being coated with acquisition
Antigen or the solid phase carrier of antibody take time long, 37 DEG C of incubations need 2-3 hours, and 4 DEG C are incubated needs and stay overnight, and adopt
It is incubated with 37 DEG C and is easily caused antigen or antibody, enzymatic activity decline, reduces detection accuracy.In addition in its specific detection process
Generally require 2-3 hours or so, so a complete ELISA process needs more than 3h.With scientific worker and clinical inspection
The information content tested the workload of personnel and contacted is increasing, and it is just extremely urgent effectively to complete more work using the limited time
Cut.The new capability that " a kind of graphene acceleration elisa technique " of the invention is combined using a kind of new material graphene with ELISA
The improved ELISA technology for accelerating antigen-antibody to combine, using the absorption of graphene height and high-ratio surface characteristic, in ELISA reactants
Increase the contact between antigen-antibody in system, ELISA reactions and coating process are not only accelerated, while improving the true of detection
Property and accuracy.
" a kind of graphene acceleration elisa technique " of the invention can be applied to containing various elisa techniques, including various bacteriums
Elisa technique, various fungi elisa techniques, various parasite elisa techniques, various virus ELISA technologies, various sensibiligens
Elisa technique, various biotoxin elisa techniques, veterinary drug residues of pesticides elisa technique, PCR-ELISA technologies, NSP-ELISA
The fields such as technology, Dot-ELISA technologies, TP-ELISA technologies, IMS-ELISA technologies, PPA-ELISA technologies.
ELISA is, by soluble antigen or antibody binding to the solid phase carriers such as polystyrene, to utilize antigen-antibody knot
Close the qualitative and quantitative detecting method that selectivity carries out enzyme labeled immunoassay reaction.Engvall and Perlmann is delivered within 1971
Enzyme-linked immunosorbent assay is used for the article that IgG is quantitative determined, and its general principle is：1. antigen or antibody is physical is attached to
Certain surface of solid phase carriers, and keep its immunocompetence.2. antigen or antibody connect into enzyme-labelled antigen or antibody with certain enzyme, this
Plant enzyme-labelled antigen or antibody had both retained its immunocompetence, the activity of enzyme is retained again.3. by inspection sample (determine antibody therein or
Antigen) and enzyme-labelled antigen or antibody reacted by the antigen or antibody of different step and surface of solid phase carriers, with the side of washing
Method makes the antigen antibody complex formed on solid phase carrier be separated with other materials, finally with reference to the enzyme amount on solid phase carrier with
The amount of tested substance is into certain ratio in sample, and after the substrate for adding enzyme reaction, substrate is changed into color products by enzymatic, production
The amount of thing is directly related with the amount of tested substance in sample, therefore can be according to the depth qualitative or quantitative analysis of color reaction.Due to
The catalytic efficiency of enzyme is very high, thus can greatly iodine effect so that assay method reaches very high susceptibility.
Graphene has following key property：1st, high electron mobility：Up to 200,000cm2V-1s-1：2nd, it is extremely low
Resistivity, resistivity is up to 10-6Ω cm, are the materials of resistivity minimum in the world, are also the superconductors under normal temperature；3、
Excellent mechanical performance：Young’s modulus 1.1Tpa；4th, high-ratio surface, the preferable graphene of quality is more reachable than surface
2600m2/g；5th, fabulous stability；6th, bio-compatibility；7th, adsorbable various atoms and molecule etc..Graphene scanning electricity
Mirror result is shown in accompanying drawing 1." a kind of graphene acceleration elisa technique " its principle of the invention is using graphene high-ratio surface, can inhaled
Attached various atoms and molecule, bio-compatibility this series of characteristics, the contact between increase antigen and antibody to accelerate ELISA,
Reaction time, which shortens, to be more beneficial for observing result and ensures its reliability.The action principle of graphene and antigen and antibody is shown in accompanying drawing
2。
" a kind of graphene acceleration elisa technique " of the invention is the elisa technique of improvement, is not for specific single production
Product, it is impossible to be described in detail one by one.Its core operation process has：1st, the preparation of graphene-containing reagent：Graphene is dissolved in buffering
Liquid and antigen or antibody with, be gently agitated for being completely dissolved it, it is closed to preserve.2nd, antigen or antibody containing graphene are coated with.
3rd, testing sample, quality-control product are added.4th, incubation, washing, colour developing, terminating reaction.5th, ELIASA is detected.
Application field citing of the present invention：
(1) infectious disease examination or quantitative detection：Communicable disease is relatively early to begin to use ELISA field, available for such as
The examination of the pathogen such as AIDS, virus hepatitis, syphilis.
(2) drug-resistant bacteria or virus screenings or quantitative detection：The resistance problems of antibiotic are increasingly taken seriously, and new drug is ground
Hair speed is much unable to catch up with the development of resistance, and elisa technique can provide early stage early warning and later stage for drug resistance problems
Detection.
(3) examination of food safety check or quantitative detection：Food security is increasingly paid attention to by everybody, enterovirus and cause
Germ and transgene component, sensibiligen, biotoxin, residue of veterinary drug, residues of pesticides in food safety check etc. can be using these
Invention carries out ELISA examinations or quantitative detection.
(4) animal quarantine examination or quantitative detection：Animal diseases allow poultry feeders to suffer huge economic loss, only examine
The disease that animal is suffered from is found, the loss of poultry feeders could be reduced as far as possible, so as to promote the development of animal husbandry, such as animal
Virus, animal bacteria, animal fungal, parazoon, animal prion etc. can carry out ELISA examinations or fixed using the present invention
Amount detection.
(5) other fields for being used for ELISA examinations, quantitatively detecting：As the detection of fermentation industry bacteria quantified, environmental protection are harmful thin
Bacterium quantitatively detects, agricultural pest examination or quantitative detection, agriculture fungi bacterium examination or quantitative detection etc..
Brief description of the drawings：
Fig. 1 is single-layer graphene SEM ESEM results.
Fig. 2 is the schematic diagram that graphene acts on antigen-antibody.Antibody is represented, ● antigen is represented,Represent not
Graphene-containing ELISA reaction systems,Represent the ELISA reaction systems of graphene-containing.
Fig. 3 is the conventional ELISA experiments for being not added with graphene.
Fig. 4 is the ELISA experiments for adding graphene reduced time.
Embodiment：
The content that the present invention is furture elucidated for detailed description below " a kind of graphene acceleration elisa technique ", but not
It is interpreted as limitation of the present invention.Without departing from the spirit and substance of the case in the present invention, to the inventive method, condition, step
Suddenly and using done modifications or substitutions, the scope of the present invention is belonged to.
The manner of formulation of main graphene-containing reagent：
(1) preparation of antigen graphene mixture：Mainly it is made up of antigen, buffer solution, high-quality graphene.Specifically prepared
Journey is as follows：Antigen is added in buffer solution, fully mixed, it is that can be used to add appropriate high-quality graphene.
(2) preparation of antibody graphene mixture：Mainly it is made up of antibody, buffer solution, high-quality graphene.Specifically prepared
Journey is as follows：Antibody is added in buffer solution, fully mixed, it is that can be used to add appropriate high-quality graphene.
Above two preparation of reagents mode, it is only for citing is convenient described, every that graphene is added by different way
Enter in ELISA reaction systems, should be understood as the justice that should have of graphene preparation of reagents, the i.e. scope of the invention.Below
Embodiment in the second exemplified by, i.e., antibody graphene mixture prepare.
Specific embodiment one：Rat c reactive protein ELISA is detected
C reactive protein (C-regctive protein, CRP) refers to be infected in body or during tissue damage in blood plasma
Some protein steeply risen (acute protein), CRP can be played conditioning and be made with the phagocytosis of activating complement and reinforcement phagocyte
With so as to remove pathogenic microorganism and the damage of invasion body, necrosis, the histocyte of apoptosis, in the innate immunity of body
Important protective effect is played in journey.The present invention detects the antigen of c reactive protein from double-antibody method, by taking 96 orifice plates as an example.
Take appropriate high-quality graphene to be first dissolved in buffer solution, fully mix, it is that can be used to add appropriate high-quality graphene,
Wherein buffer：
Above material fully dissolves, and adjusts pH to 9.6, mends ultra-pure water to 1000ml.AgainAppropriate graphene is added, is prepared
Into the buffer solution containing graphene.
The preparation of antibody graphene mixture：
The preparation of conventional antibody mixture：
Appropriate antibody graphene mixture is taken to add in each shrinkage pool of ELISA Plate (also can be other solid phase carriers) 37 DEG C
It is incubated after 2h without 4 DEG C overnight；Appropriate conventional antibody mixture is taken to add ELISA Plate (also can be other solid phase carriers) simultaneously
37 DEG C are incubated after 2h in each shrinkage pool, and 4 DEG C overnight.With confining liquid to 37 DEG C of coated ELISA Plate (also can be other solid phase carriers)
2h is closed, and the μ l of washing lotion 350 are injected per hole using artificial board-washing or machine board-washing, 1min, board-washing 5 times is soaked.Setting standard
Sample wells and sample aperture, standard sample wells respectively add the μ l of standard items 50 of various concentrations, and sample aperture first adds the μ l of sample to be tested 10, then adds dilution
The μ l of liquid 40, blank well is not added with.In addition to blank well, horseradish peroxidase (HRP) mark is added in standard sample wells and sample aperture per hole
The μ l of detection antigen 1 00 of note, reacting hole is sealed with shrouding film, and 37 DEG C of water-baths or insulating box incubate 30min.Liquid is discarded, is inhaled
Patted dry on water paper, cleaning solution is filled it up with per hole, stood 1min, get rid of and patted dry on cleaning solution, blotting paper, so repeat board-washing 5 times (
Can be machine-washed plate with board-washing).TMB chromogenic substrates and each 50 μ l of oxidant are added per hole, 37 DEG C of lucifuges are incubated 20min.Added per hole
Terminate liquid (2M H2SO4) the interior OD values that each hole is determined at 450nm wavelength of 50 μ l, 15min.
Analysis of experimental results：Using concentration known antigen as plasmid standards for quantitation, 2 times are diluted successively as standard curve, with
Standard concentration makees ordinate, and correspondence OD values make abscissa, draw out standard items linear regression curves, calculate each by curvilinear equation
Concentration of specimens value.
The conventional ELISA experiments (accompanying drawing 3) for being not added with graphene are set.Its coating time is：37 DEG C are incubated after 2h, 4 DEG C of mistakes
Night；Its reaction time is：37 DEG C of incubation 2h.Its standard concentration and corresponding OD values are as follows, and its formula is：Y=32.909x-
2.3476 R2=0.997.
| Con | 0pg/ml | 3pg/ml | 6pg/ml | 12pg/ml | 24pg/ml | 48pg/ml |
| OD | 0.005 | 0.153 | 0.245 | 0.497 | 0.791 | 1.518 |
Sample aperture OD values are 0.264, and its concentration C RP is calculated for 6.340pg/ml according to formula
The shortening time is set to add the ELISA experiments (accompanying drawing 4) of graphene.Its coating time is：Direct 37 DEG C of incubations 1h；
Its reaction time is：37 DEG C of incubation 30min.Its standard concentration and corresponding OD values are as follows, and its formula is：Y=33.223x-
2.2189 R2=0.9962.
| Con | 0pg/ml | 3pg/ml | 6pg/ml | 12pg/ml | 24pg/ml | 48pg/ml |
| OD | 0.037 | 0.149 | 0.24 | 0.493 | 0.787 | 1.494 |
Sample aperture OD values are 0.259, and its concentration C RP is calculated for 6.386pg/ml according to formula
Accompanying drawing 3,4 is analyzed：Add graphene and shorten incubation time and after the coating time, its result is routinely not added with positive
The ELISA experiments of graphene have no notable difference.In summary, in the case where other conditions are constant, adding graphene can be bright
It is aobvious to accelerate ELISA reactions.
Claims (10)
1. a kind of graphene accelerates elisa technique, it is characterized in that a kind of new material graphene adds with the ELISA new capabilities being combined
The improved ELISA technology that fast antigen-antibody is combined, using the absorption of graphene height and high-ratio surface characteristic, in ELISA reaction systems
Contact between middle increase antigen-antibody, accelerates ELISA (enzyme linked immunosorbent assay (ELISA)) reactions and coating process.
2. a kind of graphene according to claim 1 accelerates elisa technique, it is characterised in that described new material graphite
Alkene narrow sense concept refers to the only one of which carbon atom thickness for being made up of hexangle type honeycomb lattice structure with sp2 hybridized orbits carbon atom
Two dimensional crystal, can also use broad sense graphene (multilayer) to be preferred especially with the multi-layer graphene within 10 layers, and with higher
Absorption and the graphene of high-ratio surface characteristic.
3. a kind of graphene according to claim 1 accelerates elisa technique, it is characterised in that its English of described ELISA
Enzyme Linked Immunosorbent Assay are expressed as, its Chinese is expressed as enzyme linked immunosorbent assay (ELISA), and referring to can
The antigen of dissolubility or antibody binding utilize antigen-antibody binding specificity to carry out immune response on the solid phase carriers such as polystyrene
The qualitative and process that quantitatively detects.
4. a kind of graphene according to claim 1 accelerates elisa technique, it is characterised in that described graphene with
New capability that ELISA is combined accelerates the improved ELISA technology that antigen-antibody is combined, can before, during and after ELISA processes root
Appropriate graphene is added according to specific substance characteristics, to shorten the ELISA reaction time, when solving conventional ELISA one-time detections
Between long (needing 2~3 hours) and coating overlong time (needing to stay overnight) the problem of.
5. a kind of graphene according to claim 1 accelerates elisa technique, it is characterised in that described antigen be one
The material of specific immune response can be caused in organism by planting, and the material, which enters, can stimulate body to produce antibody and draw after body
Rise cellular immunity, in immunoassays antigen refer to can be with antibody binding material；It is characterized in that described antibody is a kind of
The large-scale Y for being used for differentiating and neutralizing foreign substance such as bacterium, virus etc. by immune system secreted by thick liquid cell (effect B cell)
Shape protein.
6. a kind of graphene according to claim 1 accelerates elisa technique, it is characterised in that described antigen-antibody knot
Conjunction refers to the characteristic that antigen with antibody there is selectivity to be combined, i.e. its corresponding antibody of antigen has specificity structure, by very
Weak short distance gravitation and combine, the adhesion is a kind of non-covalent bond effect power of high degree of specificity, can be to a certain degree
It is upper to be specifically bound.
7. a kind of graphene according to claim 1 accelerates the contact between elisa technique, described increase antigen-antibody
Refer to adsorb by graphene, disperse antigen and (or) antibody molecule, to increase the contact probability between antigen and antibody, and then
The joint efficiency of antigen and antibody is improved, can not only accelerate to be incubated (reaction) process, can also accelerate coating process, above-mentioned strategy is not
But it can be used for conventional ELISA, also available for fluorescence elisa technique, chemiluminescence elisa technique, PCR-ELISA skills
Art, NSP-ELISA technologies, Dot-ELISA technologies, TP-ELISA technologies, IMS-ELISA technologies, PPA-ELISA technologies etc..
8. a kind of graphene according to claim 1 accelerates elisa technique, it is characterised in that described graphene with
Contact probability this strategy that ELISA is combined between increase antigen-antibody can greatly reduce ELISA reactions and coating time, fit
For herding, agricultural, food security, clinical diagnosis, scientific research detection and enzyme immune detection and the field of immunology of diagnosis, also fit
For the immune related reagent in external diagnosis reagent II classes and III products, apply also for detecting the antigen in other samples, resist
Body and haptens etc..
9. a kind of graphene according to claim 1 accelerates elisa technique, it is characterised in that pass through a kind of green wood work stone
The absorption of black alkene and high-ratio surface characteristic accelerate ELISA strategy to be applied to computer come the contact increased between antigen and antibody
Software program and network program are write, and improve ELISA detection efficiencies, reduction ELISA reactions and coating time, further improve stone
Immunological technique of the black alkene with the ELISA acceleration ELISA processes being combined.
10. a kind of graphene according to claim 1 accelerates elisa technique, it is characterised in that pass through a kind of green wood work stone
The absorption of black alkene and high-ratio surface characteristic accelerate ELISA strategy to be applied to enzyme mark come the contact increased between antigen and antibody
Instrument, fluorescence microplate reader, spectrophotometer, sepectrophotofluorometer, automatic enzyme non-analysis meter etc. provide the instrument of detection absorbance
Device design and develop with improve ELISA reaction and coated efficiency, its main component include coating plate, enzymic-labelled antibody and (or)
Antigen, standard items, buffer solution, nitrite ion, terminate liquid, cleaning solution etc..
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710077713.1A CN107064487A (en) | 2017-02-14 | 2017-02-14 | A kind of graphene accelerates elisa technique |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710077713.1A CN107064487A (en) | 2017-02-14 | 2017-02-14 | A kind of graphene accelerates elisa technique |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107064487A true CN107064487A (en) | 2017-08-18 |
Family
ID=59599261
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710077713.1A Pending CN107064487A (en) | 2017-02-14 | 2017-02-14 | A kind of graphene accelerates elisa technique |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107064487A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113447646A (en) * | 2021-06-22 | 2021-09-28 | 浙江理工大学 | By using C-SiO2Method for accelerating ELISA detection process by using adsorbing material |
| CN115902208A (en) * | 2022-09-07 | 2023-04-04 | 深圳前海石墨烯产业有限公司 | Graphene film loaded polystyrene ELISA plate and preparation method thereof |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS62170850A (en) * | 1986-01-23 | 1987-07-27 | Wako Pure Chem Ind Ltd | Immunochemical measurement method |
| CN102226779A (en) * | 2011-03-28 | 2011-10-26 | 中国人民解放军第三军医大学第三附属医院 | Electrochemical immunodetection method |
| CN102495207A (en) * | 2011-12-26 | 2012-06-13 | 华东交通大学 | High-sensitivity enzyme-linked immunoassay method |
| CN103913573A (en) * | 2013-01-05 | 2014-07-09 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Double signal amplification ELISA detection method based on nanometer gold and graphene oxide |
| CN104132936A (en) * | 2013-12-25 | 2014-11-05 | 常州碳宇纳米科技有限公司 | Stable and novel developing solution used for determination of peroxidase |
| CN104977194A (en) * | 2014-04-10 | 2015-10-14 | 北京雷根生物技术有限公司 | Method of accelerating sample treatment with addition of graphene |
| CN105606827A (en) * | 2016-03-14 | 2016-05-25 | 山东大学 | Kit for high-sensitivity detection on proteoglycan and preparation method of kit |
| CN106124770A (en) * | 2016-06-14 | 2016-11-16 | 同济大学 | A kind of recyclable nano biological sensor of high sensitivity and preparation method and application |
-
2017
- 2017-02-14 CN CN201710077713.1A patent/CN107064487A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS62170850A (en) * | 1986-01-23 | 1987-07-27 | Wako Pure Chem Ind Ltd | Immunochemical measurement method |
| CN102226779A (en) * | 2011-03-28 | 2011-10-26 | 中国人民解放军第三军医大学第三附属医院 | Electrochemical immunodetection method |
| CN102495207A (en) * | 2011-12-26 | 2012-06-13 | 华东交通大学 | High-sensitivity enzyme-linked immunoassay method |
| CN103913573A (en) * | 2013-01-05 | 2014-07-09 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Double signal amplification ELISA detection method based on nanometer gold and graphene oxide |
| CN104132936A (en) * | 2013-12-25 | 2014-11-05 | 常州碳宇纳米科技有限公司 | Stable and novel developing solution used for determination of peroxidase |
| CN104977194A (en) * | 2014-04-10 | 2015-10-14 | 北京雷根生物技术有限公司 | Method of accelerating sample treatment with addition of graphene |
| CN105606827A (en) * | 2016-03-14 | 2016-05-25 | 山东大学 | Kit for high-sensitivity detection on proteoglycan and preparation method of kit |
| CN106124770A (en) * | 2016-06-14 | 2016-11-16 | 同济大学 | A kind of recyclable nano biological sensor of high sensitivity and preparation method and application |
Non-Patent Citations (1)
| Title |
|---|
| SANDEEP KUMARVASHIST: "Graphene-based immunoassay for human lipocalin-2", 《ANALYTICAL BIOCHEMISTRY》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113447646A (en) * | 2021-06-22 | 2021-09-28 | 浙江理工大学 | By using C-SiO2Method for accelerating ELISA detection process by using adsorbing material |
| CN113447646B (en) * | 2021-06-22 | 2024-03-19 | 浙江理工大学 | By using C-SiO 2 Method for accelerating ELISA detection process by adsorption material |
| CN115902208A (en) * | 2022-09-07 | 2023-04-04 | 深圳前海石墨烯产业有限公司 | Graphene film loaded polystyrene ELISA plate and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Clark | Immunosorbent assays in plant pathology | |
| US10234470B2 (en) | Enzyme-linked immunosorbent assay kit for detecting dinitolmide and use thereof | |
| CN101413955B (en) | ELISA test box for detecting zearalenone and preparing and detecting method thereof | |
| CN102675463B (en) | Carbendazim monoclonal antibody, preparation method and application thereof | |
| Zhu et al. | Preparation of specific monoclonal antibodies (MAbs) against heavy metals: MAbs that recognize chelated cadmium ions | |
| CN109897023B (en) | Lomitrelline hapten and artificial antigen as well as preparation method and application thereof | |
| CN103439503B (en) | The enzyme linked immunological kit of Sparfloxacin and establishment thereof and detection method | |
| CN104357401B (en) | Hybridoma, monoclonal antibody and the application of anti-II types dengue virus NS 1 monoclonal antibody can be secreted | |
| CN104928258A (en) | Canine parvovirus hybridoma, monoclonal antibody and application | |
| CN108344869A (en) | A kind of hepatitis B surface antigen chemiluminescence immune detection reagent kit and its application | |
| CN103383393B (en) | A kind of quality-control product of alternative patient's positive blood | |
| CN113637081A (en) | Hybridoma cell strain secreting pendimethalin-resistant monoclonal antibody and application thereof | |
| CN104974256B (en) | A kind of anti-thiacloprid monoclonal antibody and application thereof | |
| CN107064487A (en) | A kind of graphene accelerates elisa technique | |
| Lin et al. | Collocation of avian and mammal antibodies to develop a rapid and sensitive diagnostic tool for Russell's Vipers Snakebite | |
| CN101936997A (en) | Human anti-rabies virus IgG antibody ELISA test kit | |
| Zhu et al. | Characterization of monoclonal antibodies for lead− chelate complexes: applications in antibody-based assays | |
| CN106872693A (en) | Micro- beam preparation method and the micro- beam immune sensing detecting system based on antibody fragment modification of antibody fragment modification | |
| Liu et al. | Evaluation of a water-soluble adjuvant for the development of monoclonal antibodies against small-molecule compounds | |
| CN113416255A (en) | Anti-fasciola gigantica Cat L1 monoclonal antibody and preparation method and application thereof | |
| CN111763658A (en) | Hybridoma cell strain secreting anti-dinitrotolamine monoclonal antibody and application thereof | |
| CN104897651B (en) | A kind of chemical luminescence reagent kit of Aflatoxins M1 and its application | |
| CN104650236A (en) | Heterogenous conjugate and application thereof to RV (rubella virus) assay | |
| CN104833797A (en) | ELISA method of detecting bispecific antibody MSBODY and application of the method | |
| CN105567643B (en) | Hybridoma cell capable of secreting anti-EV 71 virus protein VP1 monoclonal antibody, monoclonal antibody and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170818 |
|
| WD01 | Invention patent application deemed withdrawn after publication |